Original Article http://dx.doi.org/10.1590/1678-77572016-0196 Microbiological, lipid and immunological SUR¿OHVLQFKLOGUHQZLWKJLQJLYLWLVDQG type 1 diabetes mellitus Abstract Cristiane DUQUE1 Objective: The aim of this study was to compare the prevalence of SHULRGRQWDOSDWKRJHQVV\VWHPLFLQÀDPPDWRU\PHGLDWRUVDQGOLSLGSUR¿OHVLQ Mariana Ferreira Dib JOÃO2 type 1 diabetes children (DM) with those observed in children without diabetes Gabriela Alessandra da Cruz (NDM), both with gingivitis. Material and methods: Twenty-four DM children Galhardo CAMARGO3 and twenty-seven NDM controls were evaluated. The periodontal status, 3 Gláucia Schuindt TEIXEIRA JO\FHPLF DQG OLSLG SUR¿OHV ZHUH GHWHUPLQHG IRU ERWK JURXSV 6XEJLQJLYDO Thamiris Santana MACHADO3 samples of periodontal sites were collected to determine the prevalence of Rebeca de Souza AZEVEDO3 SHULRGRQWDOPLFURRUJDQLVPVE\3&5%ORRGVDPSOHVZHUHFROOHFWHGIRU,/ǃ Flávia Sammartino MARIANO2 TNF-D and IL-6 analysis using ELISA kits. Results: Periodontal conditions of DM Natália Helena COLOMBO1 and NDM patients were similar, without statistical differences in periodontal indices. When considering patients with gingivitis, all lipid parameters Natália Leal VIZOTO2 evaluated were highest in the DM group; Capnocytophaga sputigena and Renata de Oliveira Capnocytophaga ochracea were more prevalent in the periodontal sites of DM 2 MATTOS-GRANER children. “Red complex” bacteria were detected in few sites of DM and NDM groups. Fusobacterium nucleatum and Campylobacter rectus were frequently IRXQGLQERWKJURXSV6LPLODUOHYHOVRI,/ǃ71)D and IL-6 were detected LQ'0DQG1'0FKLOGUHQ&RQFOXVLRQ&OLQLFDODQGLPPXQRORJLFDOSUR¿OHVDUH similar between DM and NDM children. The presence of Capnocytophaga sputigena and Capnocytophaga ochracea were associated with gingivitis in DM children. Keywords: Gingivitis. Children. Diabetes mellitus. Polymerase chain reaction. Cytokines. Submitted: May 28, 2016 0RGL¿FDWLRQ6HSW Accepted: October 27, 2016 Corresponding address: 1Universidade Estadual Paulista, Faculdade de Odontologia de Araçatuba, Departamento de Cristiane Duque Odontologia Infantil e Social, Araçatuba, São Paulo, Brasil Universidade Estadual Paulista 2Universidade de Campinas, Faculdade de odontologia de Piracicaba, Departamento de Diagnóstico Faculdade de Odontologia de Araçatuba Departmento de Odontologia Infantil e Social Oral, Piracicaba, São Paulo, Brasil. 3 Rua José Bonifácio, 1193 - 16015-050 Universidade Federal Fluminense, Faculdade de Odontologia de Nova Friburgo, Nova Friburgo, Rio Araçatuba - São Paulo - Brazil de Janeiro, Brasil. Phone: +55.18.36363315- Fax: +55.18.36363236 e-mail: [email protected] [email protected] J Appl Oral Sci. 217 2017;25(2):217-26 0LFURELRORJLFDOOLSLGDQGLPPXQRORJLFDOSUR¿OHVLQFKLOGUHQZLWKJLQJLYLWLVDQGW\SHGLDEHWHVPHOOLWXV Introduction ,/ǃ,/DQGRWKHUF\WRNLQHV29. These mediators are responsible for periodontal breakdown, leading 17 Periodontal disease comprises a group of conditions to the clinical signs and symptoms of disease . A that affects the gingiva, periodontal ligament, few studies evaluated the production of cytokines in 29 cementum, alveolar bone, and tissue structures children with gingivitis. Ulker, et al. (2008) found a that support the teeth. The predominant form of FRUUHODWLRQEHWZHHQKLJKOHYHOVRI,/ǃDQG71)ĮLQ periodontal disease in children and adolescents is JLQJLYDOFUHYLFXODUÀXLGRIFKLOGUHQDQGFOLQLFDOVLJQV gingivitis13. There is no clear-cut age at which the of gingivitis. gingival reaction to bacterial insult in children converts 6\VWHPLFLQÀDPPDWRU\GLVHDVHVVXFKDVGLDEHWHV to that found in adults. However, there is a gradual alter the host environment, and are predicted to increase in gingival activity from early childhood to increase the patient’s vulnerability to gingivitis due adult age11. WRFKDQJHVLQWKHLQÀDPPDWRU\UHVSRQVHWRPLFURELDO 12 The etiology of periodontal disease is complex. challenges . Clinical studies have demonstrated that Some bacterial species are recognized as putative the presence of diabetes can be considered a risk factor 19,31 periodontal pathogens26. In particular, Tannerella for periodontal disease in childhood . Individuals forsythia (Tannerella forsythensis), Porphyromonas with diabetes mellitus have impaired neutrophil and gingivalis and Treponema denticola, known as macrophage functioning, altered collagen production, 12 “red complex” pathogens, have been indicated and exaggerated collagenase activity , perhaps for playing important roles in various forms of OHDGLQJWRWKHSDWLHQW¶VKHLJKWHQHGLQÀDPPDWRU\VWDWH periodontal diseases7,26. Campylobacter sp., Prevotella as interactions with advanced glycation endproducts intermedia/Prevotella nigrescens, Fusobacterium (AGEs) have been shown to increase macrophage 12 19 sp., members of the “orange complex”, are also VHFUHWLRQRISURLQÀDPPDWRU\PHGLDWRUV . Salvi, et al. related to periodontal breakdown as the secondary (2010) IRXQGDKLJKFRQFHQWUDWLRQRI,/ǃLQSDWLHQWV group of periodontal pathogens and the “green with type 1 diabetes when compared to healthy 24 complex,” represented by the combination of individuals. Snell-Bergeon, et al. (2010) evaluated Eikenella corrodens, Capnocytophaga sputigena, C. 553 patients with type 1 diabetes mellitus and 215 ochracea, C. gingivalis, C. concisus was considered healthy patients aged between 10 and 22 years and primary colonizers and compatible with periodontal observed that high levels of IL-6 and other biomarkers health26. Cortelli, et al.6 (2009) detected high levels were associated with the lipid profile and may of Campylobacter rectus associated with periodontal collaborate with systemic complications in individuals health and Prevotella intermedia with the presence with diabetes. These complications could increase the 31 RILQÀDPPDWLRQ5RWLPLHWDO18 (2010) showed that, patient’s risk to develop severe periodontal disease . except for P. gingivalis, periodontopathogens such as There are few studies that have examined microbial A. actinomycetemcomitans, T. forsythia, P. intermedia colonization, immunological factors and gingival and P. nigrescens DUH UHODWLYHO\ FRPPRQ ¿QGLQJV health during childhood. The aim of this study was in the oral cavity of children. Thus, the relationship to compare the prevalence of periodontal pathogens, between clinical parameters and the prevalence of V\VWHPLFLQÀDPPDWRU\PHGLDWRUVDQGOLSLGSUR¿OHVLQ several periodontal pathogens in children need to be type 1 diabetes children (DM) with those observed in studied in greater detail. children without diabetes (NDM), both with gingivitis. The pathogenesis of periodontal disease has been widely revised16,17 and there is a consensus that, Material and methods DOWKRXJKEDFWHULDDUHHVVHQWLDOWKH\DUHLQVXI¿FLHQW for the disease to occur16. Microbial challenge in the Study population subgingival plaque modulates the host immune- The study protocol was approved by the Ethics LQÀDPPDWRU\ UHVSRQVH LQ WKH SHULRGRQWDO WLVVXHV17. Committee of Antonio Pedro University Hospital Macrophages and polymorphonuclear leukocytes, in (protocol 057/2010). Children with type 1 diabetes response to the chemo-attractant effect of bacterial mellitus (DM) and children without diabetes (NDM), toxins, such as lipopolysaccharide (LPS), are activated aged between 7 and 13 years, with mixed dentition, WRSURGXFHLPSRUWDQWLQÀDPPDWRU\PHGLDWRUV71)D, of both genders, and without distinction of race J Appl Oral Sci. 218 2017;25(2):217-26 DUQUE C, JOÃO MFD, CAMARGO GACG, TEIXEIRA GS, MACHADO TS, AZEVEDO RS, MARIANO FS, COLOMBO NH, VIZOTO NL, MATTOS-GRANER RO were selected for this study. Individuals without to the depth of the gingival sulci for 60 seconds. diabetes were recruited from the Pediatric Dentistry This procedure was performed for each of the four Clinic and children with diabetes from the database sites previously selected (mesiobuccal sulci of three kindly provided by a local Diabetes Association. The permanent molars and one permanent incisor, selected diagnosis of diabetes was given by an endocrinologist. randomly or mesiobuccal sulci of four deciduous The exclusion criteria used for subject recruitment2 molars) and the paper points of each subject were were: antibiotic prophylaxis for dental treatment, inserted in a microtube containing 1 mL of Tris-EDTA uncontrolled systemic diseases, immunological solution (10 mM Tris–HCl, 0.1 mM EDTA, pH 8.0) compromise, subjects who were wearing orthodontic RQLFH3RROHGELR¿OPVZHUHVHSDUDWHGDFFRUGLQJWR devices, subjects who had been undergoing periodontal dentition (permanent or deciduous) for each patient. treatment 12 months before the beginning of the 7KHVDPSOHVZHUHVWRUHGDWí&XQWLOWKHDQDO\VHV study, those who had been taking antibiotics within 6 months prior to the clinical examination, those with Blood samples collection extensive caries lesions, individuals who were using Patients were asked to reduce the intake of fatty an antiseptic solution during 3 months period and foods the night before collecting the blood samples. smokers. Parents or legal guardians were informed of Blood samples were collected by a specialized the study and signed an informed consent form and professional from the peripherical vein (cubital fossa) completed an interview regarding the medical and of individuals who had an overnight fast. Samples dental histories of the children. were collected in vacuum collection tubes and sent to Raul Sertã Hospital Laboratory at Nova Friburgo/ Clinical measurements RJ for clinical analysis [fasting glucose levels (GL), The following clinical parameters were measured: